Fig 2: M1, M2 macrophages contribute to the different steps of retinal neovascularizaion (RNV). RNV at P13 in the retinas of mice with OIR. Retinas were stained with isolectin B4 and CD11c, and at P18 retinas were stained with isolectin B4 and CD206. Flatmounts were examined by laser scanning confocal microscopy. Isolectin B4-positive endothelial tip cells and macrophages are shown in (A–C). Yellow indicates the co-localization of endothelial cells and CD11c-positive cells [M1 macrophages (D–F)] or CD206-positive cells [M2 macrophages (G–I)]. Costaining of CD11c with F4/80 at P13 is shown in (J–L) and CD206 with F4/80 at P18 in (M–O). CD11c-positive cells interact with endothelial tip cells at the vascular front, while CD206-positive cells embrace the emerging vessels and bridge the neighboring vessel sprouts, suggesting a promotive function for tip cell fusion (n=4 mice/group). OIR, oxygen-induced retinopathy; NOR, normal; P, post-natal day.

Fig 3: ITGAX (CD11c) gene expression and CD11c plaque area are associated with a vulnerable plaque phenotype and symptomatic carotid plaques in humans. (A) Using an OPLS-DA analysis, IRF5 and ITGAX (CD11c) were identified as the myeloid cell surface marker and transcription factor and with the largest impact on separating symptomatic (<31 days prior to surgery) plaques from asymptomatic plaques. RNAseq data from 47 human carotid plaques (24 symptomatic and 23 asymptomatic). Blue indicates asymptomatic and red indicates symptomatic. (B) Human plaque gene expression levels of IRF5 and ITGAX (CD11c) were also significantly increased in plaques associated with symptoms within 31 days prior to surgery compared with asymptomatic plaques. Lines indicate mean levels, each dot represents an individual value (n = 60, 34 symptomatic and 26 asymptomatic). (C) CD11c+ but not IRF5+ plaque area, as assessed by immunohistochemistry, was increased in symptomatic carotid plaques. Lines indicate median levels, each dot represents an individual value (n = 62, 35 symptomatic <31 days prior to surgery and 27 asymptomatic). Data are presented as % of total plaque area. (D) OPLS-DA analysis identified ITGAX as the myeloid cell gene with the largest impact on separating plaques with a vulnerability index above median from plaques with a vulnerability index below median (n = 47, 24 symptomatic and 23 asymptomatic). Blue indicates less than median vulnerability index and red indicates greater than median vulnerability index. (E) IRF5 and CD11c plaque areas are increased in plaques with a high (above median, n = 31) vulnerability index compared with plaques with a low vulnerability index (below median, n = 31). Mann–Whitney U tests were used. (F) Plaque area stained positive for CD11c and IRF5 correlated with the calculated vulnerability index (n = 62). Spearman test was used for the correlation analysis. CD11c, cluster of differentiation 11c; IRF5, interferon regulatory factor 5.

Fig 4: Effects of Paenalcaligenes hominis and Escherichia coli on the occurrence of cognitive impairment and colitis in specific pathogen-free mice. Effects on the occurrence of cognitive impairment in Y-maze (A), NOR (B), and Barnes maze tasks (C). D Effects on the BDNF expression and NF-?B activation in the hippocampus. Effects on the BDNF+/NeuN+ (E), NF-?B+/Iba1+ (F), TLR4+/Iba1+ (G), LPS+/Iba1+ (H), and IL-1R+ cell populations (I) into the hippocampus. J Effects on the IL-1ß expression in the hippocampus. K Effects on the endotoxin levels in the blood, assessed by LAL assay kit. Effects on the colon length (L), myeloperoxidase (MPO) activity (M), IL-1ß expression (N), and NF-?B+/CD11c+ cell population (O) in the colon. P Effects on the LPS levels in the feces. Paenalcaligenes hominis (PH, 1 × 109 CFU/mouse/day) and Escherichia coli (EC, 1 × 109 CFU/mouse/day) were orally gavaged for 5 days. Control mice (NC) were treated with vehicle (saline) instead of bacterial suspension. Data values were indicated as mean ± SD (n = 6). Means with the same letters are not significantly different (p < 0.05). A, B, C, K, L, M, N Kruskal-Wallis test with Dunn’s post hoc test for non-parametric analysis. J, P One-way ANOVA with post-hoc Bonferroni’s multiple comparisons test

Fig 5: miR-19b overexpression results in repressed p-STAT3 and CD11c levels in AC eye and CLN tissues. (A) Expression levels of miR-19b in conjunctiva, cornea, and CLNs derived from the PBS, AC, AC + CYT387, AC + miR-19b, and AC + control miR groups as determined by qPCR. The results are expressed as mean ± SD from three independent experiments and each experiment was performed in triplicate. (B) Immunofluorescent staining on frozen sections of eyeballs from mice in each group that were probed for p-STAT3 (red) and CD11c (green) with DAPI counterstaining (blue). The scale bars represented 100 µm. *P < 0.05, **P < 0.01, ***P < 0.001, and ****P < 0.0001. The experiments were repeated 3 times.